Enhancing GvL via delayed ex-vivo co-stimulated DLI after non-myeloablative SCT

项目描述（由申请人提供）：本项目的长期目标是建立一个新的临床平台，通过预防性输注体外共刺激的同种异体DLI来增强移植后的移植物抗白血病（GvL）效果，以改善急性白血病和骨髓增生异常综合征患者的预后。该策略基于我们的假设，即在供体嵌合建立后，在最小残留病（MRD）的时候，在移植后早期没有炎症环境的情况下，激活的供体t细胞将诱导更有效的GvL效应，而不会引起严重的GvHD。本研究将为实施肿瘤特异性过继细胞治疗，即在不增加GvHD的情况下增强GvL提供基础。具体目标1：开展I期临床试验，以确认高风险恶性血液病成人患者在t细胞耗尽非清髓性异基因干细胞移植后延迟输注活化DLI的可行性和安全性。移植物抗宿主病的发生率和严重程度将是主要终点。特异性目标2：评估预防性激活DLI对肿瘤特异性细胞毒性和免疫重建的影响：2a)研究t细胞对白血病特异性抗原（自体肿瘤）、多克隆刺激（SEB、PMA）和召回抗原（CMV、EBV）的反应，通过基于3个互补读出的供体t细胞功能功能分析，评估在体外共刺激前后供体t细胞的免疫潜力增强：(i)通过CFSE流式细胞术检测细胞增殖，（ii）通过干扰素γ释放（ELISPOT）检测细胞因子分泌，（iii）通过CD107a流式细胞术检测细胞毒性，作为特异性靶杀伤的标志物。2b)研究t细胞对白血病特异性抗原（自体肿瘤）、多克隆刺激（SEB、PMA）和召回抗原（CMV、EBV）的增殖反应（增殖、细胞因子分泌、细胞毒性），通过使用上述5个时间点对受体t细胞进行功能分析，评估输注离体共刺激供体细胞前后受体t细胞的免疫重建增强；移植前，大约+90天（pADLI #1前），大约+160天（pADLI #2前），+270天和+365天。

DESCRIPTION (provided by applicant): The long-term goal of this project is to establish a novel clinical platform for enhancing immune reconstitution, and more specifically, the graft vs leukemia (GvL) effect post-transplant via prophylactic infusion of ex-vivo co-stimulated allogeneic DLI to improve the outcome for patients with acute leukemia and myelodysplastic syndrome. This strategy is based on our hypothesis that activated donor T-cells given after donor chimerism is established, at a time of minimal residual disease (MRD), and in the absence of the inflammatory milieu of the early post-transplant period, will induce a more potent GvL effect without causing severe GvHD. This study will provide a foundation for efforts to implement tumor-specific adoptive cellular therapy, i.e., enhancing GvL without increasing GvHD. Specific Aim 1: Conduct a phase I clinical trial to confirm the feasibility and safety of delayed infusion of activated DLI after T-cell depleted non-myeloablative allogeneic stem cell transplantation in adult patients with high risk hematologic malignancies. The incidence and severity of graft vs host disease will be a primary endpoint. Specific Aim 2: Assess the impact of prophylactic activated DLI on tumor-specific cytotoxicity and immune reconstitution: 2a) Study T-cell responses to leukemia-specific antigen (autologous tumor), polyclonal stimuli (SEB, PMA), and recall antigens (CMV, EBV), to assess for the enhanced immune potential of donor T-cells before and after ex-vivo costimulation via functional analysis of donor T-cell function based on 3 complementary read-outs: (i) proliferation via CFSE flow cytometric assay, (ii) cytokine secretion via interferon gamma release (ELISPOT), and (iii) cytotoxicity via degranulation assay as a marker of specific target killing based on flow cytometric analysis of CD107a. 2b) Study T-cell proliferative responses (proliferation, cytokine secretion, cytotoxicity) to leukemia-specific antigen (autologous tumor), polyclonal stimuli (SEB, PMA), and recall antigens (CMV, EBV), to assess for enhanced immune reconstitution of recipient T-cells before and after infusions of ex-vivo costimulated donor cells via functional analysis of recipient T-cells using the assays outlined above at 5 time points; pre- transplant, approximately day+90 (pre pADLI #1), approximately day+160 (pre pADLI #2), day+270, and day+365.